N-isopropylacrylamide-co-2-hydroxypropyl methacrylate) hydrogels were synthesized using free radical initiated copolymerization method. Four hydrogels with different cross-linker concentrations were prepared. Semi-crystalline, cross-linked copolymer networks were confirmed by FTIR, SEM and XRD analysis. Variation of swelling behaviour was monitored gravimetrically and thermo-responsiveness has been noticed. An application of synthesized thermo-responsive hydrogels as carriers for the modulated release of anti-inflammatory model drug was investigated. Moreover, naproxen loading into these hydrogels was also determined using FTIR, SEM and XRD techniques and release was analyzed using HPLC method at simulated physiological conditions. Swelling kinetic and mechanism of water transport, as well as diffusion of naproxen through the hydrogels were analyzed. Thus, the aim of this work was to study various compositions of obtained hydrogels and their possibility of application as a thermo-responsive carrier for prolonged naproxen release in order to evaluate as a potential candidate for drug carrier in future pharmaceutical applications.In this study, poly( Nonsteroidal anti-inflammatory drugs (NSAIDs) are generally used in the symptomatic treatment of rheumatoid arthritis, osteoarthritis, inflammation, acute gout, dysmenorrhoea, metastatic bone pain, headaches, migraines and postoperative pain. They do not cause tolerance and dependence, but the characteristic of their application is the phenomenon of reaching a plateau, i.e., maximum effect that cannot be exceeded by increasing the dose. Most of these drugs exert three types of effects: anti-inflammatory, analgesic and antipyretic effect . NaproxeIn order to achieve effective targeted delivery and satisfactory therapeutic effects of naproxen, different polymers were designed as drug carriers. Numerous of investigations outlining ways to improve low solubility of naproxen were published. In the inclusion complexes with β-cyclodextrins, naproxen solubility and release rate were increased ,8. Ilić-Stimuli sensitive polymers are intensively studied for preparation of controlled drug delivery systems for variety of active pharmaceutical ingredients ,12. NaprN-isopropylacrylamide), (NiPAm) and its properties were applied to design a series of thermo-sensitive polymeric systems. Interpenetrating polymer networks with N-isopropylacrylamide/acrylamide were obtained via in situ free-radical polymerization using synthesized macro-cross-linkers poly(ethylene glycol diacrylate) and poly(ε-caprolactone diacrylate) [N-isopropylacrylamide by a series of polymerization reactions. Poly(MAH-β-cyclodextrin-co-NIPAAm) hydrogels demonstrated slow naproxen release efficiency [N-isopropylacrylamide-co-methacrylamide) compared to the poly(N-isopropylacrylamide) showed that release from the copolymer was more sustained than that from the homopolymer. [N-isopropylacrylamide and hydroxypropyl methylcellulose thermosensitive hydrogel for the naproxen sodium release was investigated [N-isopropylacrylamide-co-poly(g-glutamic acid)-allyl glycidyl ether) hydrogels were obtained by cross-linking polymerization and the morphology, thermal-and pH sensitivity and biodegradability were studied. The hydrogels were loaded with the naproxen and the release studies were performed under simulated physiological conditions [Stimuli-responsive hydrogels are materials that can be designed for pharmaceutical application and have an important role in modulated/controlled drug release. They are inert and compatible with the human body tissues and some internal organs . They hecrylate) . The resficiency . Propertpolymer. . The infstigated ,32,33,34nditions .N-isopropylacrylamide-co-2-hydroxypropyl methacrylate), p(NiPAm-co-HPMet), co-hydrogels, as drug carriers for modulated release of naproxen. The object of the presented study was to provide a naproxen release formulation with modified characteristics resulting in continued levels of the drug above a 24 h period by using thermosensitive properties of synthesized hydrogels. In previous papers, the potential applications of hydrogels for controlled release of caffeine [NiPAm-co-HPMet) copolymer indicated that the copolymer could be used as an appropriate carrier for modified drug delivery.The goals of presented study are application, characterization and evaluation of a series of copolymer poly 99%, 2-hydroxypropyl methacrylate (HPMet) 96.5% and 2,2′-azobis(2-methylpropionitrile) (AZDN) 98% by Acros Organics, Morris Plains, NJ, US; ethylene glycol dimethacylate (EGDM) 97%, methanol Chromasolv for HPLC, 99.9% HPLC grade and naproxen 98.5–100.5% by Fluka Chemical Corp., Buchs, Switzerland; potassium bromide for IR spectroscopy, Potassium bromide (KBr), ≤100%, by Merck KGaA, Darmstadt, DE; acetone by Centrohem, Belgrade, RS; methanol, p.a. by Unichem, Belgrade, RS.N-isopropylacrylamide with 15 mol% of co-monomer 2-hydroxypropyl methacrylate, in relation to the amount of NiPAm monomer, were prepared using EGDM for cross-linking and 2.8 mol% of AZDN for the first step of initiation by free radical polymerization method. The cross-linker EGDM concentrations of 0.5, 1.0, 1.5, 2.0 and 3.0 mol% in relation to the total co-monomers mass in the reaction mixtures were employed to observe the effect of cross linker on co-hydrogels. After all reactants diluted in acetone, the homogenized reaction mixture was syringed into glass ampoules, which were then sealed using the heat of the flame while the ampoule was rotating. Sealed ampoules were heated in the temperature regime: 120 min at 70 °C, 60 min at 80 °C, and 30 min at 85 °C until all initiator’s bonds were homolytically cleaved, producing radicals. After cooling, the obtained co-hydrogels were separated from the glass ampoules and cut into smaller cylinders . Then, for the purpose of removal of all non-reacted compounds, hydrogels were extracted by methanol during 72 h, after that they were submerged into methanol/distilled water solutions in 75/25, 50/50, 25/75 and 0/100% for a day. Swollen co-hydrogels were dried to constant mass at 40 °C to pass into the xerogel stage for further testing.Cross-linked copolymers consisted of monomer tα, and equilibrium swelling ratio, α∞, were according to Equations (1) and (2), respectively:m0 is the xerogel mass, tm is the mass of the swollen hydrogel at the time t and m∞ is the mass of the swollen hydrogel at equilibrium state.Swelling of co-hydrogels was monitored gravimetrically at the temperature of 5 °C and 38 °C. Four synthesized samples in xerogels states were weighed and then immersed in fluids with simulated physiological pH values 7.4 and 2.0. At predetermined and regular time intervals, the hydrogels were taken out, wiped with filter paper to remove excess fluid, mass were recorded and returned back in the same vial until constant mass was achieved. The swelling ratio, NiPAm-co-HPMet) xerogels were swollen at equilibrium state in distilled water at 5 °C, weighed again and then immersed into the same vials during the temperature increasing to 60 °C. At each particular temperature, hydrogel samples were incubated in distilled water to the equilibrium state for 24 h, wiped with filter paper, weighed tm and m∞, and then were put back. The swelling ratio and equilibrium swelling ratio were evaluated using Equations (1) and (2).The temperature responsiveness of co-hydrogels was monitored within the temperature range of 5–60 °C in a temperature controlled water bath . The weighted p.In order to analyze the nature of the fluid diffusion process into the co-hydrogels with different cross-linker contents, Fick’s Equation (3) was applied to fit and calculate the experimental data ,40.(3)Mn and k were determined from the slope and intercept of the linear relationship between lnF and lnt. The diffusion coefficient D was calculated for the initial phase of swelling to 60% from Equation (5) [l is the thickness of the dried sample (cm), and D is the diffusion coefficient (cm2/min). By taking the logarithm of the Equation (5), the Equation (6) is obtained (the linear relationship between ln(t/MeM) and lnt):D can be calculated from the intercept of the linear relationship between ln(tM/eM) and lnt, where t is the time for which the gel absorbs half the total amount of fluid. n, which determined the mechanism of water or solvent diffusion [By taking the logarithm of the Equation (3), the Equation (4) is obtained:tion (5) ,42,43: polymeric network, synthesized xerogels were swollen in the naproxen standard substance solution, 40 mg/cm3 in methanol/distilled water mixture, 80/20, during 48 h at temperature of 5 °C. Naproxen incorporation efficiency (η) was calculated using Equation (7):gL is the mass of naproxen incorporated into the co-hydrogel (mg/gxerogel) and uL is the maximum of naproxen available mass for incorporation (mg/gxerogel).In order to study the possibilities of the application of synthesized co-hydrogels as carriers for the modified release of nonsteroidal anti-inflammatory drugs, the naproxen was applied as a model drug. For the purpose of naproxen incorporation into the p. The amount of the released naproxen was monitored for 24 h. Aliquots (100 µL) from the medium with released naproxen were taken in certain intervals of time , diluted with methanol, filtered through a 0.45 μm cellulose membrane filter and analyzed using HPLC method. The quantification of the amount of the released naproxen as a function of time was performed using the optimized and validated method developed by Somia and colleagues [® STAR C18, 25 cm × 4.6 mm, 5 µm  were applied. The mobile phase was methanol:water, 90:10 (v/v), at pH 2.7 adjusted with phosphoric acid, at the flow rate 1.2 cm3/min with isocratic elution. Agilent ChemStation software was used to process the obtained chromatograms.The swollen co-hydrogels with loaded naproxen were soaked with 7 cmlleagues . HPLC AgNiPAm-co-HPMet) xerogels, and p(NiPAm-co-HPMet) with incorporated naproxen were grounded in the Amalgamator  to reduce the particle size to less than 5 mm in diameter. FTIR spectra of powder samples  xerogels, and p(NiPAm-co-HPMet) with incorporated naproxen) with the KBr (about 0.6% related to KBr amount) were subjected to vacuuming and pressing under a pressure of about 200 MPa to form transparent KBr pelets. All FTIR spectra were recorded on a Bomem Hartmann & Braun MB-series FTIR spectrophotometer  in the wavelength range of 4000–400 cm−1. Win-Bomem Easy software was applied for FTIR spectra analysis.Powder samples of synthesized p. The swollen hydrogels were firstly frozen and after that the amount of fluid was removed by sublimation at −30 °C and 0.05 kPa during 18 h, and in the isothermal desorption phase they were heated at 20 °C at the same pressure. Freeze-dried co-hydrogels were packed under vacuum and stored at 5–7 °C.NiPAm-co-HPMet) copolymers in xerogels state and freeze-dried hydrogels in equilibrium swollen state, before and after naproxen incorporation were first metalized with a gold/palladium (85/15) alloy under vacuum using JEOL Fine Coat JFC 1100E Ion Sputter . After coating, they were scanned on a JEOL Scanning Electron Microscope JSM-5300 .Samples of p(NiPAm-co-HPMet) xerogels, naproxen and xerogels with incorporated naproxen were analyzed on Philips PW1030 powder diffractometer  by being exposed to monochromatic Cu Kα radiation in the range of 2θ = 7–34° with 0.05° increments and recording time of τ = 5 s. The operating device voltage and the strength of the electric current were 40 kV and 20 mA, respectively.Powder samples of synthesized p band and sI is the peak intensity of the secondary band. CrI is an empirical measure of relative crystallinity. The variation of the crystallinity index (∆CrI) can be determined using the Equation (9):N-co-HPMetiPAm and CrINiPAm are values of crystallinity index for copolymer p(NiPAm-co-HPMet) and homopolymer p(NiPAm), respectively.The crystallinity index (CrI) was determined using the empirical models to evaluate the degree of crystallinity developed by Segal and co-workers ,46, and p values < 0.05 were considered to be statistically significant.The swelling study, XRD and in vitro naproxen release study were carried out in triplicate. Standard deviation of mean of triplicate data from the individual data was quantified using Origin software, for statistical analysis. The statistical analysis was carried out by one-way analysis of variance (ANOVA) and NiPAM-co-HPMet) hydrogels was performed with 15 mol% of co-monomer HPMet and 0.5, 1.0, 1.5, 2.0 and 3.0 mol% of EGDM. The obtained product with 0.5 mol% of EGDM cross-linker remained in liquid consistency after polymerization without required hydrogel consistency, and it was not used for further analysis.The synthesis of p(NiPAm-co-HPMet) xerogel samples during swelling in distilled water was monitored at range of 5 °C–60 °C. The swelling ratio, αt, was calculated according to Equation (3), and the equilibrium swelling ratio, α∞, according to Equation (4). The mass of xerogel samples (m°) was measured until equilibrium was established (m∞).The behaviour of p(NiPAm-co-HPMet) hydrogels with different cross-linker content  at different pH values (2.0 and 7.4) and temperatures (5 °C and 38 °C)   was moniThe swelling capacity of a series of co-hydrogels depends on the temperature of swelling medium and gradually decreases when the cross-linker amount increases . All analyzed samples in distilled water increasingly swelled from the initial moment to the first 400 min of swelling at both temperatures.α∞ = 29.07 for sample with lesser cross-linker amount (1.0 mol% of EGDM) (α∞ = 21.46) [At 5 °C co-hydrogels swelled gradually until equilibrium swelling was reached after 2 days at of EGDM) a. It can= 21.46) . This loα∞ = 4.04 hydrogel with 1.0 mol% of EGDM) presented in previous papers [The co-hydrogels swell significantly lesser at 38 °C and reach equilibrium state after 400 min, i.e., 1 g of hydrogel with 1 mol% EGDM can absorb nearly 4.56 g of water from the sample with 1 mol% of EGDM b. The acs papers ,48. In bs papers ,37.The analysis of obtained results showed that the swelling ratio decreases with increasing EGDM content, due to many available cross-linker’s molecules that form a polymer network with a lot of cross-linking places. The small hole free volume in copolymer network with a higher cross-linker content reduces the possibility of swelling and an equilibrium swelling ratio was achieved in a shorter time. In contrast, when the EGDM cross-linker’s amount is lesser, long-chain structures have fewer cross-linking places at the same distance, causing the polymer network pore to expand more and thus increase the water absorption capacity. According to free volume theory, the total free volume could be divided into interstitial free volume (which is not implicated in facilitating transport through the liquid) and the hole-free volume, and self-diffusivity of penetrant could be easily predicted . It is pNiPAm-co-HPMet) with 17% of HPMet obtained with an applied γ-ray dose of 20 kGy in water has a swelling rate of about 5.5. A similar result with equilibrium swelling ratio of about 5.0 was reached for hydrogel p(NiPAm-co-HPMet) with 10 wt% of HPMet, obtained by radical polymerization in aqueous solution with 2% of N,N-methylene-bis(acrylamide) as a cross-linker [The achieved swelling ratio for co-hydrogels synthesized by gamma radiation method have a lesser value compared to hydrogels obtained by free radical polymerization . The cros-linker .α∞) and swelling kinetic parameters for synthesized co-hydrogels: diffusion exponent (n), kinetic constant proportionality (k) diffusion coefficient (D), and the correlation coefficient (R2), determined by using Equations (3)–(6), at 5 °C and 38 °C at pH values 7.4 and 2.0 are shown in Values of equilibrium swelling ratio  for all samples, i.e., the solvent transport in the polymeric network is considerably slower than the relaxation of polymer chains [NiPAm-co-HPMet) at 25 °C have been reported in the literature [k, are higher at 38 °C due to the reduction of the hole free volume in the polymer network [D, at 5 °C are in range of 0.326–5.379 × 10−6 cm2/min, while at 38 °C, above lower critical solution temperature (LCST), they are in the range of 7.066 × 10−5–1.899 × 10−4 cm2/min, indicating faster diffusion, which is similar to the results published in the literature [The values of the diffusion exponents, r chains . The pror chains ,53,54. Oterature ,38. The  network . The valterature ,56,57.NiPAm-co-HPMet) hydrogels, was monitored throughout the change in the swelling ratio at the temperature increasing from 5 °C to 60 °C in distilled water in order to test their temperature responsivity.The influence of temperature to the swelling properties of synthesized p(NiPAm-co-HPMet) with 1.0 mol% EGDM, and the sample with 3 mol% EGDM showed a very small change in the degree of swelling and phase transition. The highest values of swelling capacity for all samples of co-hydrogels were achieved at 5 °C, when e.g., 1 g of a sample with 1.0 mol% of EGDM absorbed 29.08 g of water. As the temperature increased to 60 °C, the hydrogels dehydrated with the release of free, unbound water from the polymer network, so that 1 g of the same sample retained only 2.06 g of water. This behavior of hydrogels showed similarity with the results previously published in the literature [All analyzed co-hydrogels show a “smart” reaction which is reflected in a significant decrease in the swelling ratio with increasing external temperature, which can be seen in terature ,38,47,58terature ,60. ThisNiPAm, synthesized p(NiPAm-co-HPMet) hydrogels with 1.0, 1.5, 2.0 and 3.0 mol% of EGDM are presented in NiPAm-co-HPMet) with the FTIR spectrum of NiPAm monomer, some differences can be noticed, which indicates formation of new structures, i.e., some bands are not present in the spectrum of NiPAm monomer, absence of several bands characteristic for monomers, as well as the change of intensity and position of certain bands. In all FTIR spectra of the p(NiPAm-co-HPMet) there are no absorption bands at 1620–1640 cm−1, which originate from the stretching vibration of C = C double bond and confirm that the polymerization reaction is performed via breaking double bonds of vinyl groups from monomers and cross-linkers. The intensity of the bands originating from the stretching vibrations of vinyl group = C-H (maximum in the monomer FTIR spectrum at 3071 cm−1) significantly decrease and shift by 5 units to lower wavenumbers, confirming that polymerization occurred by breaking the double bond of vinyl groups [NiPAM trapped into the polymer network can be noticed. In all FTIR spectra of copolymers, a broad bands in the range from 3600 cm−1 to 3100 cm−1 were observed, where the first bands at 3311 cm−1 originate from the stretching vibrations of side N-H bonds, ν(NH), with maxima shifted by 10 units to lower wavenumbers than in the spectrum of monomer. Second band maxima at 3436 cm−1 were the results of the stretching vibrations of O-H bonds, ν(OH), from co-monomer HPMet present in the side-chains of polymer network. In the p(NiPAm-co-HPMet) spectra the maxima intensities of amide bands I, ν (C=O), were shifted by 4 units to higher wavenumbers (1652 cm−1) and amide bands II, δ(N-H), of amide bands II were shifted by 2 unit to lower wavenumbers (1554 cm−1), indicating that C=O and N-H bonds participated in the formation of intramolecular hydrogen bonds. The presence of the bands originating from stretching vibrations of C-H bond of methyl and methylene groups in the range of 2800 to 3000 cm −1, as well as the stretching vibrations of keto groups C=O from ester, ν (C=O), at 1729 cm−1 were noticed in all spectra. Individually divergences in absorption bands intensity in the FTIR spectra of series p(NiPAm-co-HPMet) copolymers compared to the monomer NiPAM are a consequence of the polymerization and crosslinking reaction during the formation of hydrogel. FTIR spectra a series of synthesized p(NiPAm-co-HPMet) copolymers are matching with the published data [FTIR spectra of monomer l groups . The prehed data .NiPAm-co-HPMet), sample with 1.0 mol% of EGDM, with incorporated naproxen and naproxen are shown in NiPAm-co-HPMet) copolymer. Since free -OH groups, amide groups and ester groups are present in the structure of this hydrogels, they may be important for non-covalent interactions with naproxen, especially with the ether, C-O and carboxyl groups, i.e., via the carbonyl C=O and -OH group. For this reason, changes in the maxima position of these functional groups’ bands were monitored. The band of stretching vibrations of the -OH group from the p(NiPAm-co-HPMet) in the spectrum of this copolymer with incorporated naproxen exists and it is shifted by 3 units to lower values of wavenumbers (3433 cm−1), which means that it participated in the formation of non-covalent interaction. Additionally, maxima displacements of the bands from the naproxen‘s -OH group, from in-plane and out-of-plane bending vibrations were observed in the spectrum of copolymers with naproxen. The in-plane bending vibrations, δ(OH) absorbs at 1389 cm−1, are shifted by 5 units, while out-of-plane bending vibrations δ(OH) are shifted by 1 unit (925 cm−1) both towards lower wavenumbers, which confirms their participation in the interaction with the copolymer. Additionally, the shifts in the stretching vibrations band of the C=O group from the ester (1729 cm−1), towards higher wavenumbers for 1 unit relative to the spectra of the copolymers and for 2 units relative to the spectra of naproxen were observed. The N-H group stretching vibration band from the copolymer side-chains in the spectrum of copolymers with naproxen was shifted by 19 units (3292 cm-1) to lower wave numbers also indicating the interaction with naproxen. The maximum of amide band I in the spectrum of copolymers with naproxen occurs at 1650 cm−1 and is shifted by 2 units to lower wavenumbers of copolymers, while the maximum of amide band II is shifted by 8 units to higher wavenumbers and occurs at 1552 cm−1. The band of stretching vibrations of C-O group from the ether‘s part of the naproxen structure was shifted by 3 units to higher wavenumbers in the spectrum of copolymers with naproxen, occurring at 1230 cm−1, indicating that it interacted with free -NH or secondary -OH groups from side chains of copolymers by the hydrogen bond formation. Analysis of the FTIR spectra indicates that hydrogen bonds were formed between the naproxen and the copolymers via the carboxyl group, i.e., -OH and C=O and ether C-O groups of naproxen and -OH and -NH groups in the copolymer side chains.The FTIR spectra of p(NiPAm-co-HPMet) copolymers with 1.0, 2.0 and 3.0 mol% of EGDM obtained by X-ray diffraction analyses were presentetd in Three XRD diffractograms samples of synthesized p(NiPAm-co-HPMet) copolymers samples contain two broad, unstructured diffraction peaks with low intensity and indicate a low degree of crystallinity, a small crystallite size and dislocations of the crystal structure at large distances. The first peaks (s) are narrower and occur at diffraction angle values around 8°. The second, broader peaks, named fundamental (f), occur at a diffraction angle of about 20°, which is consistent with published data [NiPAM) homopolymer, synthesized using free radical polymerization method, analog as described, which was identical as previously published [NiPAm-co-HPMet) presented a high similarity with XRD pattern of homopolymer p(NiPAM). The value of diffraction intensity for fundamental (If) and secondary (Is) peaks from and diffraction angle values of p(NiPAm-co-HPMet) and p(NiPAM) hydrogels were presented in All of three XRD patterns of p hydrogel sample with 1.0 mol% of EGDM after naproxen incorporation were also presented in XRD diffractograms of naproxen and p(f) and secondary (Is) peaks from XRD patterns and diffraction angle values for the co-hydrogel after naproxen incorporation were presented in In the XRD diffractogram of co-hydrogel with incorporated naproxen  absence NiPAm-co-HPMet) samples show two maxima of the reflection. In the samples with reduced EGDM content the intensity of secondary peaks (Is) increases (NiPAm-co-HPMet) reduces crystallinity and intensifies the amorphousness in comparison to XRD diffractogram of p(NiPAM) homopolymer (Diffraction lines of p(ncreases . This isopolymer .NiPAm-co-HPMet) copolymers before and after naproxen incorporation. The secondary (Is) peak was shifted in the diffractogram of co-hydrogel with incorporated naproxen. This is probably result of hydrogen bonds formed between the naproxen and the co-hydrogel, and it is in correlation with analysis of FTIR spectra.X-ray diffraction analysis is most frequently used to measure the crystallinity in the polymers. Two simple empirical models to evaluate relative crystallinity of co-hydrogels were applied. The X-ray crystallinity index (CrI) was determined by the Equation (8) and the variation in the crystallinity index (∆CrI) was calculated by the Equation (9). The obtained values of CrI and ∆CrI for the synthesized co-hydrogels samples are given in NiPAm-co-HPMet) copolymers in xerogel state were given in SEM micrographs in Morphology of synthesized p(NiPAm-co-HPMet) hydrogels reveals porous nature of hydrogel network. The presence of pores in the co-hydrogel structure facilitates the water passing through the hydrogel network, which is especially important in hydrogels application as drug carriers. It can be estimated that the pores in the xerogel state are on average about 30 μm and bigger than 70 μm in swollen state. Additionally, the alternation of semi-ordered/crystalline and amorphous regions are observed, which is probably due to the influence of the amount of cross-linker on the size and distribution of pores. Cracks of up to more than 5 μm are also observed as well as the appearance of more regulated areas, which also reflected on the swelling ability.The SEM micrographs of synthesized p(NiPAm-co-HPMet) with incorporated naproxen is displayed by SEM micrographs in The surface appearance of the copolymer p(NiPAm-co-HPMet) with incorporated naproxen  are given in NiPAm-co-HPMet) is in the range 56.82–87.02%. There is a trend of decreasing efficiency of fitting as the hydrogel cross-linking density increases and the share of co-monomer HPMet increases. Synthesized porous hydrogel network allows the absorption of relatively large drug quantities.The efficiency of naproxen incorporation into p(NiPAm-co-HPMet) hydrogels which is schematically presented in By analyzing the results obtained by the FTIR, XRD and SEM methods, it can be concluded that naproxen was successfully incorporated into the p(t = 5.821 min.The content of released naproxen from co-hydrogels was determined based on the Equation (8), which is valid for the linear part of the constructed calibration curve. The appearance of the peaks on the HPLC chromatogram originating from naproxen is shown in max = 206 nm and originates from the allowed π→π* transition of the disubstituted naphthalene, and the second is more intense at λmax = 233 nm and originates from n→π* transitions due to the presence of the keto group, C=O, with the transition at λmax = 256 nm, due to the conjugation of substituted propionic acid [−8 μg/cm3. The limit of determination, (LOQ) as the lowest amount of a substance that can be reliably  determined by an analytical method, is LOQ = 3.62 × 10−8 μg/cm3 [The characteristic UV spectrum of naproxen b shows tnic acid . The lim8 μg/cm3 .The results of the in vitro cumulative release of naproxen from the synthesized co-hydrogels at temperatures at 38 °C in fluid with pH 7.4 and pH 2.0, which simulate human body temperature and pH are shown in NiPAM-co-HPMet) sample with the lowest cross-linker content (1.0 mol% EGDM) released the highest amount of naproxen . The smallest amount of naproxen was released from sample with 3 mol% of EGDM , in accordance with the expectation and results of the swelling. During the first 24 h about 50% of naproxen remained inside the hydrogels, and this amount would be appropriate for prolonged release.The results show higher values of released naproxen at a temperature above LCST. At 38 °C, in fluid pH 7.4 during 24 h p, and the least from the sample with the highest EGDM cross-linker content  during 24 h (At 38 °C in acidic fluid pH 2.0 the most naproxen was released from the co-hydrogel sample with 1.0 mol% of EGDM (119.93 mging 24 h a. The amNIPAM-co-HPMet) does not have an ionic functional group, low sensitivity to changes in the pH value of fluid was expected.Temperature responsiveness of co-hydrogels was dominant in aggregation process and naproxen releasing. They contract and squeeze fluid with naproxen when temperature increase above VPTT. The applied amount of naproxen had not significant influence on the aggregation process. At temperatures above the LCST dominated hydrophobic interactions lead to a decrease in the hole free volume, which causes the broken intermolecular hydrogen bonds with naproxen and active drug release due to hydrogel contraction. The obtained results show a lower amount of released naproxen in acidic fluid than alkaline fluid. The reason is maybe the lower solubility of naproxen in acidic fluid. Additionally, it can be attributed to the effect of propionic acid dissociation from naproxen structure. As the molecular structure of p of the naproxen release mechanism from hydrogels were assessed by fitting experimental release using Equations (3)–(6) and are presented in The kinetic parameters (NiPAm-co-HPMet) hydrogels with incorporated naproxen could be suitable as a carrier for modulated of NSAID release.Kinetic parameters of the naproxen release from the co-hydrogels show that the mechanism of fluid transport belongs to Fickian, i.e., “Less Fickian” diffusion . The druNiPAm-co-HPMet) copolymer by FTIR spectrum confirms the success of copolymerization process of NIPAM with HPMet comonomers. XRD difractograms and SEM micrographs verified semi-crystalline structure of obtained hydrogels. The swelling transport mechanism of the co-hydrogels at 5 °C and 38 °C in alkaline fluid corresponds to “Less Fickian” diffusion for all samples, i.e., the solvent transport in the polymeric network is considerably slower than the relaxation of polymer chains. One sample at 5 °C and pH 7.4 and all samples in acidic fluid showed anomaly in the mechanism of water transport, i.e., the swelling process is controlled by water diffusion and relaxation of polymer chains. The structural changes resulting from the incorporation of the naproxen as a model drug into the p(NiPAm-co-HPMet) copolymers determined by FTIR analysis indicate that non-covalent type intermolecular interactions between the copolymers and the incorporated naproxen were created. The surface structures of p(NiPAm-co-HPMet) in xerogels and hydrogels state, indicate the incorporation of the naproxen into the copolymer pores. The efficacy of naproxen incorporation ranged in proportion to the cross-linker content. The formulation of p(NiPAM-co-HPMet) hydrogels with incorporated naproxen in this investigation may be used as a potentially efficient drug carrier for prolonged naproxen release when body temperature increases.The structural analysis of the synthesized p(http://www.zis.gov.rs/home.59.html. https://worldwide.espacenet.com/patent/search/family/046025219/publication/RS53220B?q=RS53220B.Registered patent RS53220B: Ilić-Stojanović, S.; Nikolić, Lj.; Nikolić, V.; Petrović, S.D.; Stanković M. Process for synthesis of thermosensitive hydrogels and pharmaceutical applications, The Intellectual Property Office of the Republic of Serbia,


N = 26; 13 from each condition) were offered the B4DT. None of the patients declined participation, and there were no dropouts. At post-treatment 59.5% were in remission, 31.0% had treatment response, and 9.5% showed no change. At 3-month follow-up 71.4% were in remission, 19.0% had treatment response, and 9.5% showed no change. There were also significant improvement in self-reported symptoms of OCD, generalized anxiety symptoms, and depressive symptoms. The results are in line with what we previously have found for the B4DT in a number of effectiveness studies. In addition, the results indicate that the patients who previously had received an unsuccessful SH intervention and patients who had first been in a WL condition, profited as much as patients who had received the B4DT as the initial intervention. Implications for clinical guidelines and for dissemination of the B4DT are discussed.The Bergen 4-day treatment (B4DT) for obsessive-compulsive disorder (OCD) was recently tested in a randomized controlled trial, where the results showed that the B4DT was more effective than a self-help intervention (SH) and a waiting list condition (WL). Patients in the SH and WL condition still in need of treatment after the first intervention ( Obsessive-compulsive disorder (OCD) is a disabling disorder characterized by recurrent, unwanted thoughts, images, or urges (obsessions) and/or repetitive behaviors or thoughts (compulsions) . CognitiSince 2012, the OCD-team in Bergen, Norway, has developed a highly concentrated exposure based treatment format for patients with OCD, the Bergen 4-day treatment (B4DT). During four consecutive days, OCD patients receive individually tailored and therapist assisted exposure treatment delivered in a group setting, where the patient–therapist ratio is 1:1. The approach is highly attractive to the patients, and there is basically no attrition. A number of open trials indicate that more than 90% of the patients can expect to respond to treatment with nearly 70% being recovered 1–4 years after participating in the B4DT , 2019. TRecently, the first randomized controlled trial (RCT) of the B4DT was completed , where iThe patients who were in need of further treatment after the SH or WL condition were offered the B4DT, which implies that for the patients who first received the SH, the National Institute for Health and Care Excellence  guidelinFrom 2016 to 2017 a randomized controlled trial (RCT) was conducted at Solvang Community Psychiatric Service  in Norway . The RCTThe inclusion criteria for the study were: (a) signed informed consent, (b) ≥18 years, (c) met DSM-5 criteria for OCD, (d) Yale-Brown Obsessive Compulsive Scale (Y-BOCS) score ≥16, and (e) spoke Norwegian fluently. The exclusion criteria for the study were: (a) meeting DSM-5 criteria for bipolar disorder, psychosis, or ongoing substance abuse/dependence, (b) intellectual disability based on previous medical history, (c) an eating disorder in need of medical attention, (d) not willing to refrain from anxiolytics during treatment, (e) ongoing suicide ideation, (f) SSRI-treatment initiated or dosage changed during the last four weeks before starting treatment, and (g) living far away from the treatment center (>1.5 h). Patients with prior CBT for OCD were not included due to a nationwide trial targeting this population. The patients who were not helped after a SH condition or a WL were offered the B4DT . A flow-n = 16) function as a comparison condition at follow-up. The 26 patients who had received either WL or SH condition were assigned to the first available B4DT group.Two of the patients who had received the SH intervention had improved according to their own evaluations and did not want further treatment and one patient who had received SH moved to another part of the country. Thus, 13 patients in the previous SH condition started treatment with the B4DT. Of the 15 patients who previously completed the WL condition, one moved to another part of the country, and another received inpatient treatment due to pregnancy complications, leaving 13 patients who started the B4DT. All patients who had received B4DT as the immediate intervention  = 0.121, p = 0.731. In addition, there were no significant differences between these two groups on any other pre-treatment variable. All patients but one were classified as having severe OCD (Y-BOCS score of 24 or more) at pre-treatment.The patients’ use of medication was recorded at the initial interview. Patients using benzodiazepines discontinued these prior to treatment. Compliance was confirmed in the post-treatment assessment. Patients had no other treatment during the four days, as confirmed by medical records.Patients had a diagnostic-  and Y-BOThe Y-BOCS , b is a Two self-report measures of OCD symptoms were also included. The Dimensional Obsessive Compulsive Scale Short-Form DOCS-SF;  is a fivThe second self-report measure of OCD symptoms was the Obsessive-Compulsive Inventory-Revised OCI-R; , an 18-iIn adition to measures of OCD-symptoms, the study included measures of comorbid symptoms of depression and generalized anxiety. The Patient Health Questionnaire-9 PHQ-9;  is a 9-iThe Generalized Anxiety Disorder Scale GAD-7;  is a 7-iIn addition to the symptom scales, we also included a measure of satisfaction with treatment. The Client Satisfaction Questionnaire 8 CSQ-8;  is an 8-There were equal number of patients and therapists in each group, and each group was led by an expert on the treatment format. All therapists were trained in delivering the treatment . Prior tA detailed description of the treatment format is given in Prior to exposure treatment, patients are taught to pay close attention to the way the exposures are done, and one of the main features of the 4-day treatment, namely the “LEan into The anxiety” (LET-technique) is demonstrated. Patients have to approach stimuli that elicit anxiety/discomfort and act in a manner that is incompatible with their OCD. During the two days of exposure treatment, the patients are assisted when they practice the LET-technique, and encouraged and assisted to employ the approach in all situations that elicit anxiety/discomfort.Participants receive psychoeducation in which they are provided with explanations for the treatment and for the techniques that are seen as essential. The psychoeducation is manualized using a PowerPoint presentation with a text-file detailing the content to be presented. The patients share their suggested exposure tasks with the group, and individual plans are made together with input from the group leader.Throughout Day 2 and 3, patients practice individually tailored and therapist assisted ERP. In the evening patients are to continue with self-administrated training. Exposure sessions are interspaced with brief group meetings where each patient share their ratings of their own exposure performance with the group. They evaluate their own LET-performance on a 1–6 scale. A score of 6 indicates that they were “leaning fully into” the anxiety. Scores below 6 are explored by the group leader, and the patients are given advice and encouragement aimed at giving them courage to “not hold back” in the next exposure. Patients were given the opportunity to invite relatives and friends to a psychoeducation session in the afternoon of day 3.Day 4 is used for summarizing and planning self-exposures for the next three weeks. Patients are taught to be their own therapists. They make their own exposure plans to be performed the first three weeks after the treatment. Written information concerning procedures and upcoming assessments is handed out, and the group ends with the patients providing feedback on their experiences with treatment.Patients log homework (exposures) completed for a 3-week period after treatment. About 12 weeks after treatment, patients meet for an individual follow-up session at the clinic. This session is intended to help the patient summarize their experiences after completing treatment, and if necessary, to refresh the treatment principles. This follow-up session did not involve exposure exercises.An adaptation of the international consensus criteria was used for assessing treatment response , not incThere were few instances of missing data. For Y-BOCS and the self-report scales there were four patients missing data at post-treatment-2. However, all four patients attended the follow-up assessment, and their scores from follow-up were therefore carried backward to replace their missing post-treatment-2 score. Imputation using “next observation carried backward” is one of the imputation methods giving least bias in scores . There wt-tests were first used to investigate if the SH or WL groups had any significant improvement on Y-BOCS scores (post-treatment-1). Paired sample t-tests were also used to investigate whether the two groups had a significant improvement after being given the B4DT at post-treatment-2 and 3-month follow-up. Independent sample’s t-tests were used to compare the two groups at post-treatment-2 and follow-up. Possible differences in change scores (from pre-treatment to post-treatment-2 and follow-up) between participants given immediate B4DT and delayed B4DT were also compared using independent samples t-tests.Paired sample Treatment outcome was further analyzed using repeated measures ANOVA with three different times of assessment . This analysis was used for all outcome measures .t-tests showed that there was no significant effect of the WL at post-treatment-1, while there was a significant but small improvement observed for the SH-condition. The patients then received the B4DT after the post-treatment-1 assessment. Irrespective of which original condition the patients belonged to they achieved a very large and significant improvement at the post-treatment-2 assessment. The change scores (pre-treatment to post-treatment 2/follow-up), was not significantly different between the conditions at post-treatment 2; t(24) = 0.164, p = 0.871, or at follow-up; t(24) = 0.35, p = 0.73. Participants from the two groups were therefore merged together into one group for further analyses.A summary of the Y-BOCS scores for the SH- and WL-conditions is displayed in N = 16) and patients originally in the SH or WL condition (N = 26). The immediate B4DT group had a mean change score of 15.85 (5.85) at post-treatment compared to 16.31 (4.52) for the SH/WL patients, t(40) = 0.287, p = 0.775. There were also a non-significant difference at follow-up, as immediate B4DT showed a mean change score of 18.19 (5.95) compared to 17.58 (7.18) for the WL/SH group, t(40) = 0.285, p = 0.777. Thus, for the analysis of clinical change all patients were combined into one group having received the B4DT, either directly or after another condition.Change in Y-BOCS scores after being given the B4DT was compared between patients given immediate B4DT  = 0.33, p = 0.74, from those not taking such medication. However, they showed tendencies toward having higher Y-BOCS scores at post-treatment  and follow-up , p = 0.048 and p = 0.066, respectively. Similar observations were made for the DOCS-SF data, but not for the other outcome measures . There were no significant differences in client satisfaction scores, t(22) = 0.09, p = 0.93.Patients using psychotropic medication did not differ on OCD-severity at pre-treatment, Data on treatment satisfaction is presented in The present study was a follow-up of a recently conducted RCT for OCD where the experimental group was treated with the newly developed B4DT and the two control conditions consisted of patients that either were allocated to SH or WL. The results from the controlled trial  showed tThe results indicate that both the patients who had previously received SH and those in the WL condition benefited significantly from the B4DT, and that they did not differ in treatment effect at 3-month follow-up from the patients who had received the B4DT immediately. At post-treatment and follow-up 90.5% showed a treatment response, and the remission rates were 58% at post-treatment and 73.1% at follow-up. This corresponds well with previous research on the B4DT e.g., . OverallThere are only three studies investigating the effect of stepped care treatment for OCD, and they suffer from sample size and attrition issues. In a pilot study of stepped care, 11 patients started with SH (step 1), followed by self-directed ERP with minimal therapist contact (step 2) if they did not respond to the first step of treatment, and step 3 was traditional therapist directed ERP . Of the The results suggest that there should be no methodological or obvious ethical concerns to first randomize OCD-patients to 3-month of WL or a SH program before they receive the B4DT. Given that patients usually have to wait before they receive treatment, this finding is interesting because it underscores that it is likely that patients can benefit from the B4DT even if they do not receive the treatment immediately. This can be of value, e.g., when patients express an urge to initiate treatment as soon as possible. Patients who had received SH did not gain more from the B4DT post-treatment or at follow-up as compared to the WL-patients. On the other hand, the patients with prior SH, were not at a disadvantage regarding the benefit they experienced following the B4DT. It is noteworthy that the SH was delivered without assistance from a therapist and that the patients were guaranteed B4DT after the SH treatment if they needed further treatment.There are some limitations of the current study such as limited sample size and lack of long-term follow-up. However, studies using the B4DT with larger sample sizes and long-term follow-up assessments have indicated that the results are consistent. Another limitation concerns the fact that the SH condition was not assisted. It is likely that assisted SH could yield better treatment results , 2015. FThe results from this RCT provide evidence that most OCD patients treated with B4DT, delivered at another site than the originators’ clinic, achieve highly comparable outcomes at post-treatment and 3-month follow-up, as a number of open trials have reported , b. In aThe datasets generated for this study are available on request to the corresponding author.ClinicalTrials.gov (Identifier: NCT02886780). Patients still in need of treatment after the SH intervention and the WL, were for ethical reasons granted participation in a B4DT group if they wanted. Also, the length of the SH and WL conditions were not longer than the ordinary waiting period at the clinic at the time when the trial was initiated. The patients/participants provided their written informed consent to participate in this study.The trial received ethical approval from the Regional Committee for Research Ethics  and was registered in GK, L-GÖ, BH, KH, and GL contributed to the study design. GL and KH contributed to the data collection. All authors contributed to the statistical analysis, interpretation of the data, and drafting of the manuscript, and approved the final version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The importance of hypoxia in the pathophysiology of inflammatory bowel disease (IBD) is increasingly being realized; also, hypoxia seems to be an important accelerator of brain inflammation, as has been reported by our group and others. IBD is a chronic intestinal disorder that leads to the development of inflammation, which is related to brain dysfunction. However, no studies have reported whether hypoxia is associated with IBD-induced neuroinflammation. Therefore, the objective of the present study was to determine whether hypoxia augments cerebral inflammation in a DSS-induced colitis mouse model. The mouse model was developed using 3% DSS for five days combined with exposure to hypoxic conditions  for two days. Mice were randomly divided into four groups: control group, DSS group, hypoxia group, and DSS plus hypoxia group. The results demonstrated that DSS combined with hypoxia resulted in up-regulation of colonic and plasmatic proinflammatory cytokines. Meanwhile, DSS plus hypoxia increased expression of Iba1, which is a marker of activated microglia, accompanied by increased expression of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in the brain. Moreover, the expression of tight junction proteins, such as zonula occludens-1 (ZO-1), occludin, and claudin-5, was markedly downregulated. The current study provides new insight into how hypoxia exposure induces excessive inflammatory responses andpathophysiological consequences in the brain in a DSS-induced colitis model. Inflammatory bowel disease (IBD), which is comprised of two main types, ulcerative colitis (UC) and Crohn’s disease (CD), is characterized by chronic or relapsing immune activation and inflammation within the gastrointestinal tract that markedly give rise to diarrhea, cramping, and pain, which are all standard symptoms of IBD -treated adult male rats exhibited a marked inflammatory response in the brain that was characterized by microglial activation and TNF-α production, leading to increased susceptibility to seizures  signaling  is through the disruption of the blood-brain barrier . In all the experiments, the ethics guidelines for investigations involving conscious animals were followed, and the experiments were approved by the Animal Care and Use Committee of the Institute of Basic Medical Sciences (NO. IACUC-2017049). Seven- to eight-week-old male C57BL/6 mice weighing 18 ± 2 g were housed under a 12-h light/dark cycle with free access to standard rodent chow and water. The mice were maintained under specific-pathogen-free (SPF) conditions.A well-characterized chemical model was used to initiate experimental IBD by giving the mice access to 3% DSS  in their drinking water for 5 days combined with exposure to hypoxia  for 2 days. The mice were divided into four groups  and sacrificed on day 7 after the beginning of DSS administration in the experiments. Colonic tissue, brain tissue, and serum were collected for the experiments.The mice were anesthetized with 1% sodium pentobarbital by intraperitoneal injection and sacrificed. Then, the colons were isolated and sectioned. The colons were washed twice in pre-cooled PBS and soaked overnight in paraformaldehyde . After immersion in absolute ethyl alcohol through a graded alcohol series, the tissues were embedded in paraffin, sectioned, and stained. Then, the colonic tissue sections were scanned by a Nanozoomer-XR Scanner C12000 .To explore the effects of hypoxia on the development of DSS-induced colitis, the mice were individually evaluated daily to determine the body weight, stool consistency, and bleeding, as detailed in Total RNA was isolated from mouse distal colon tissues using TRIzol reagent according to the manufacturer’s instructions . Reverse transcription was performed by a reverse transcription kit . cDNA was amplified by real-time PCR using SYBR Green master mix  as recommended by the manufacturer. Oligonucleotide primer sequences used for real-time PCR are in g, 30 min). TNF-α, IL-1β, and IL-6 in serum were measured using a mouse Quantikine ELISA kit  according to the manufacturer’s instructions.Blood samples were clotted for 2 h at room temperature before centrifugation  for 30 min at 37°C and then incubated with a specific primary antibody  overnight at 4°C before incubation with a secondary antibody  for 90 min at 37°C. The color reaction was developed by incubation with a 3,3′-diaminobenzidine (DAB) solution for approximately 3 min. The sections were imaged under a microscope .The mice were anesthetized with 1% sodium pentobarbital by intraperitoneal injection and perfused with prechilled saline (0.9%) to remove the circulating blood cells. The brains were washed twice with precooled PBS and soaked overnight in paraformaldehyde  after each brain was dehydrated and frozen-sectioned at thicknesses of 20 μm. First, peroxidase was removed using 10% HThe protein expression levels of ZO-1, occludin, and claudin-5 in the cortex tissues were determined. The samples were homogenized using RIPA lysis buffer on ice. Each sample was separated by SDS–PAGE. Then, the proteins were transferred to polyvinylidene difluoride membranes. After blocking, specific primary antibodies  were incubated overnight at 4°C. After washing, the membranes were incubated with HRP-conjugated secondary antibodies  for 1 h at room temperature. The specific bands were visualized using an ECL detection kit . Quantification of the band intensities was performed by normalization to β-actin.post hoc test was used for evaluating multiple comparisons. For all analyses, *p < 0.05, **p < 0.01 and ***p < 0.001 were considered statistically significant.The data were analyzed using GraphPad Prism 7.0. The data are presented as the arithmetic mean ± SEM. Statistically significant differences between groups were determined using one-way ANOVA with Dunnett’s test. Two-way ANOVA followed by Bonferroni’s ad libitum for five days. To study the effect of hypoxia on IBD, the mice were exposed to hypoxia  after DSS treatment. The mice were sacrificed and examined for colitis on day 7 : the control group, DSS group (3%), hypoxia group, and DSS+ hypoxia group. First, to evaluate brain inflammation in this model, we performed immunohistochemical staining for Iba-1 and analyzed the microglial population in the cortex and hippocampus. The microglial population in the cortex and hippocampus significantly increased after DSS + hypoxia-induced colitis . Also, tThe function of the BBB is maintained by TJs, and we assessed the TJs using western blotting. We evaluated the molecular changes in the TJs in the mouse brains. Although ZO-1, occludin, and claudin-5 did not exhibit marked changes after DSS treatment alone, ZO-1, occludin, and claudin-5 were downregulated in the mice that were subjected to DSS combined with hypoxia exposure. DSS + hypoxia treatment markedly decreased the ZO-1, occludin, and claudin-5 levels in the brain cortex compared with DSS treatment alone . These rHypoxia has long been associated with gastrointestinal dysfunction; hypoxia has also been proposed to be a risk factor for the development of neuroinflammation. However, whether hypoxia regulates neuroinflammation in colitis remains elusive. Here, we demonstrate that DSS + hypoxia resulted in abnormal expression of proinflammatory cytokines in the colon and plasma. Also, we showed increased expression of Iba-1, which is a marker of activated microglia, accompanied by increased expression of TNF-α, IL-1β, and IL-6 in the brain. Moreover, tight junction proteins were markedly downregulated.ad libitum for five days combined with exposure to hypoxic conditions for two days. In the present study, hypoxia may play a pro-inflammatory role initially in DSS-induced colitis. Hypoxia affected innate immune cells in the lamina propria of the intestine, neutrophils exhibited a delayed apoptosis rate, resulting in pathological injury induced by inflammation  was combined with recombinant IL-1β exposure, the translocation of tight junction proteins and decreased transendothelial electrical resistance (TEER) were observed .LD, YH, HL and LZ conceptualized the study. LD, XL and YG prepared and maintained the mice. YH and LD designed and performed morphological analysis and biochemical assays. MZ, TZ, XC and MF provided the materials. YH wrote the manuscript. LZ, LG and LD discussed and edited the manuscript. LZ and HL supervised the project. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The standard therapy for American cutaneous leishmaniasis (ACL) is intravenous meglumine antimoniate (IV-MA). However, treatment interruptions due to adverse events (AEs) and non-adherence are frequent. Consequently, intralesional MA (IL-MA) was proposed.This study examined the effectiveness of and AEs associated with IL-MA.2 of lesion area) once weekly for 1–8 weeks. IV-MA was performed via intravenous injections of MA at a dosage of 10–20 mg Sb5+/kg/day for 20 days. The primary outcome was defined as a lesion cure 3 months after treatment, and AEs were secondary outcomes.We performed a retrospective cohort study of 240 patients with ACL. We excluded patients with mucous lesions and disseminated leishmaniasis and those who received treatment in the previous 6 months. We considered protocol treatments as the main risk factors. IL-MA was performed using a subcutaneous injection of MA in a volume sufficient to elevate the lesion base  = 0.68, 95% CI: 0.50–0.92, p < 0.001), while the rate of AEs was similar. Female sex , lesion diameter ≤1 cm  and treatment with IV-MA  were independently associated with achieving a cure. Comorbidities  were independently associated with AEs.Patients of IL-MA group were older, had more comorbidities and more previous unsuccessful treatment of ACL. Nevertheless, IL-MA had a cure rate of 66.7%, and it was useful in this context. A prospective randomized trial is recommended.  •Intralesional antimonials (IL-MA) had a 66.7% lower cure compared to systemic.•IL-MA group was older, had more comorbidities and unsuccessful previous treatment.•IL-MA was useful in this group, where worse clinical responses are expected.•Lesions larger than 3 cm did not have more chance of failure with IL-MA.•There was no mucous recurrence after IL-MA in an area of New World Leishmaniasis. The mosyanensis . Clinicayanensis . ACL genyanensis . Treatme5+/kg/day for 20 days , administered parenterally at a dose of 10–20 mg Sb 20 days . This th 20 days . Severe  20 days . Pregnan 20 days .Treatment modalities to reduce toxicity, improve efficacy and facilitate administration and adherence have emerged for the treatment of ACL in recent decades. Injections of intralesional MA (IL-MA) have been performed in reference centres in Brazil for more than 20 years. The Pan American Health Organization (PAHO) included IL-MA as an alternative therapy via consensus in 2013 and emphasized the low level of evidence . The MinThe present study evaluated the ACL cure and AEs rates in patients who received IL-MA compared to patients who received intravenous MA (IV-MA).22.1We performed a retrospective cohort study and included ACL patients treated with MA at the University Hospital of Brasília, Brazil, from 1999 to 2017. ACL case definition relied on clinical, laboratory and epidemiological criteria described elsewhere . ACL was2.25+/kg/day or a daily maximum of 15 mL. For the IL-MA group, we used a cure rate of 56%. This rate was justified by the scarcity of existing data and a level of arbitration based on our clinical expertise. A sample calculation was performed using SAS 9.4 software considering a difference in the percentage of cure between the two groups of 34% . For theOur main register located 982 patients who could potentially fulfil the pre-defined inclusion criteria. The HUB recently started the use of an electronic register for consultation. Older patient files are still stored in paper files, and those files are kept in warehouse located 15 km from the hospital, which makes accessing them a challenging bureaucratic task. Due to complete files are difficult to access, we used a list of identification data  that allowed a first screening of 240 eligible patients, which was part of our target population. Due to these internal difficulties and to make the study feasible, we used a well-validated random sampling technique to provide a picture of the population profile (cure rate) with a 90% power at a significance level of p < 0.05 .Fig. 1Fl2.32 of lesion area), with a maximum of 5 mL, once weekly for 1–8 weeks. Alternatively, IV-MA therapy was performed using intravenous 10–20 mg Sb5+/kg/day for 20 days according to the PAHO and Ministry of Health of Brazil recommendations , treatment duration, and interruption time. Treatment interruption was considered when patients stopped treatment for over one week in the IL-MA group based on institutional protocol, and over one day in the IV-MA group or once the antimony half-life was approximately 32.8 ± 3.8 h . If this criterion was not fulfilled, the patient was considered not cured at this time, and a new treatment was performed, according to a routine protocol. The occurrence of AEs was considered a secondary outcome. We classified AEs according to their potential effect on treatment continuation: mild: patient should be closely monitored ; and severe: treatment must be interrupted immediately . According to institutional routine, patients were monitored weekly during treatment with MA to identify AEs, which were characterized as clinical, laboratory and electrocardiographic changes that occurred during treatment, with no possible causal relationship to external factors.2.55+/kg/day), group , comorbidities, interruption, cure and AEs, with no or yes answers. The determination of the age cut-off was based on recommendations from the Ministry of Health , age , duration of lesion , number of lesions , size of lesion , antimonial dose . The number and size of lesions, treatment duration, antimonial dose and other variables in the IL-MA group were not associated with a higher chance of cure or AEs. The risk of AEs was not different between groups. The classification of AEs as mild and severe was also not different . Among t3.35+/kg/day and the application of IV-MA  for up to 10 sessions . In New There was no difference in the prevalence of interruption between the groups in the present study. Most reasons for treatment discontinuation were due to AEs and rarely due to adhesion problems. The institutional protocol in this reference centre was weekly follow-up during treatment. This schedule had a positive influence on patient adherence to both treatments. We did not observe a difference in AEs prevalence or level between the groups. We observed changes in cardiac and hepatic tests during treatment with IL-MA. Antimonial intralesional absorption occurs , and it The cure rate of IV-MA in our study was 98.1%. A previous ACL meta-analysis reported a cure rate of 76.9% , and theThe subjects of this retrospective study were followed-up for at least 3 months, but 36 (69.23%) patients in the IV-MA group and 13 (61.9%) patients in the IL-MA group had a follow-up of at least 1 year. Twenty-six (50%) of the patients in the IV-MA group and 10 (47.62%) patients in the IL-MA group had a follow-up of at least 2 years. Ten (19.23%) of the patients in the IV-MA group and 2 (9.52%) patients in the IL-MA group were followed for more than 5 years. No mucosal or recurrent cutaneous leishmaniasis was reported in the patients of this study. Other studies of ACL also showed no mucosal involvement after intralesional therapy in long-term follow-up . The limThe intralesional technique had a lower cure rate than the intravenous antimonial technique and a similar AEs rate in this cohort study. However, the IV-MA and IL-MA groups had different baseline characteristics: patients treated with IL-MA were older, had more comorbidities and more previous ACL treatment attempts. Therefore, it was expected that this group would have a lower cure rate and increased risk of AEs. The IL-MA group had a cure rate of 66.7% and similar AEs as the IV-MA group, which is a good result for this special population. Nevertheless, IL-MA is a useful treatment option for elderly patients, patients with comorbidities and patients with other unsuccessful specific systemic treatment attempts. No progress to mucosal injury was observed in the patients follow-up. A prospective randomized trial with a representative sample is recommended to further evaluate this practical technique.None.


Insulin-like growth factor 1 (IGF-1) is the standard biochemical marker for the diagnosis and treatment control of acromegaly and growth hormone deficiency (GHD). However, its limitations necessitate the screening for new specific and sensitive biomarkers. The elonginB/C-cullin5-SOCS-box-complex (ECS-complex) (an intracellular five-protein complex) is stimulated by circulating growth hormone (GH) and regulates GH receptor levels through a negative feedback loop. It mediates the cells’ sensitivity for GH and therefore, represents a potent new biomarker for those diseases. In this study, individual ECS-complex proteins were measured in whole blood samples of patients with acromegaly (n = 32) or GHD (n = 12) via ELISA and compared to controls. Hierarchical clustering of the results revealed that by combining the three ECS-complex proteins suppressor of cytokine signaling 2 (SOCS2), cullin-5 and ring-box protein 2 (Rbx-2), 93% of patient samples could be separated from controls, despite many patients having a normal IGF-1 or not receiving medical treatment. SOCS2 showed the best individual diagnostic performance with an overall accuracy of 0.93, while the combination of the three proteins correctly identified all patients and controls. This resulted in perfect sensitivity and specificity for all patient groups, which demonstrates potential benefits of the ECS-complex proteins as clinical biomarkers for the diagnostics of GH-related diseases and substantiates their important role in GH metabolism. Growth hormone deficiency (GHD) and acromegaly are two growth hormone disorders caused by reduced or increased levels of growth hormone (GH), respectively. Those diseases are mainly triggered by pituitary neuroendocrine tumors (PitNETs) that slowly shift the physiological balance of hormone secretion to pathological levels. While PitNETs appear to be rather common within the general population, with 17%, only a very small percentage <0.04%) of affected people develops symptoms of a GH disorder that requires medical attention . Regardl% of affeBecause symptoms of GH-related disorders evolve very subtly over the course of many years, initial diagnoses are made only after 5–25 years . This reFor the diagnosis of GH disorders, it is suggested to gather biochemical evidence by performing at least one laboratory test . Such prThe response to medical treatment is highly individual and neither provocative tests nor biochemical markers are able to reliably predict treatment control . This maSeveral potential alternatives to IGF-1 have been proposed but despite clinical correlation with GH-related diseases in those markers, none of them got established in clinical diagnostics ,31,32,33Here, we investigated a novel set of potential biomarkers that have a close connection to the physiological action of GH. The molecule of interest is the elonginB/C-cullin5-SOCS-box-complex (ECS-complex) which consists of five proteins: Suppressor of cytokine signaling 2 (SOCS2), elongin B (EloB), elongin C, cullin-5 and ring-box protein 2 (Rbx-2). This complex controls GH signaling by regulating the GH receptor (GHR) through a negative feedback loop. As the GH uptake of target cells is primarily dependent on the GHR expression, the proteins of the ECS-complex are assumed to reflect the individual cellular response characteristics towards GH . The proThe regulatory action of the ECS-complex takes place, when GH binds to the GHR. This phosphorylates the receptor and activates the intracellular Janus Kinase/Signal Transducer and Activator of Transcription pathway, which induces the expression of SOCS2. Together with the other four proteins, SOCS2 subsequently forms the ECS-complex in the cytoplasm and is then able to bind to phosphorylated tyrosines at the GHR. This allows Rbx-2 to transfer ubiquitin moieties to the GHR, which in turn is recognized and degraded by the proteasome see . SOCS2 dThe GHR shows differential expression in peripheral blood mononuclear cells (PBMCs), which suggests that the regulatory ECS-complex is active in these cells . A GH-deA positive approval from the Ethics Committee of the Landesärztekammer Berlin  was obtained previous to starting the study and all participants gave their written informed consent before blood withdrawal. The control group participants were recruited in a local general practitioner’s surgery. They had to be above the age of 40 and were without known neoplasms or hormonal disorders. Common cold or flu were exclusion criteria, and normal IGF-1 values were verified to eliminate possible GH-related abnormalities in the control group. For the experimental groups, we included patients from four endocrine surgical departments who had been diagnosed with either acromegaly or GHD. The diagnosis of acromegaly was determined by IGF-1 and inadequate suppression of serum GH to <0.4 ng/mL by oral glucose load. GHD was diagnosed by IGF-1 and insufficient GH response to insulin-induced hypoglycemia.2/EDTA vacutainers, put immediately on ice and were transported to the laboratory within a few hours. Red blood cells were lysed with 0.17 M ammonium chloride and PBMCs collected from 2 mL of whole blood by two centrifugation cycles of 5 min at 453× g. After washing, pelleted PBMCs were frozen at −80 °C.From every subject, a single whole-blood sample of 5 mL was obtained during their routine medical consultation. Blood samples were drawn between 7 and 12 am, were stored in KIGF-1 measurements of serum from patients and controls were conducted with the Immulite 2000 (Siemens AG). For the classification of the samples, the age-dependent in-house reference range was used .6 cells per mL in PBS and homogenized using the Precellys (Bertin Instruments) and 1.4 mm zirconium oxide beads at 5.000 rpm for 15 s. The beads were removed, and the solution was centrifuged at 5.000× g for 5 min. The resulting supernatant contained the proteins of interest.For the protein measurements, PBMCs extracted from whole blood samples were thawed in cold phosphate buffered saline (PBS) and the cell concentration was determined with an automated cell counting system . Afterwards, cells were diluted to 4 × 10The levels of the ECS-complex proteins SOCS2, EloB, cullin-5 and Rbx-2 were measured with sandwich enzyme-linked immunosorbent assay kits purchased from Abbexa Ltd.  in accordance with the manufacturer’s instructions. Due to a lack of an appropriate ELISA kit or antibody, the ECS-complex component elongin C was not analyzed in this study.All data are given as mean ± standard deviation (SD) unless stated otherwise. Pearson correlation coefficients and unpaired t-tests were calculated using GraphPad Prism 5. Graphics were created with R statistics v3.5.2. P-values lower than 0.05 were considered significant. Protein concentrations have been estimated from the signal intensities of ELISA tests with the five parameter logistic regression model (5PL) of Prentice  and normion (1)) .(1)xi=ln = 14, male = 8, female = 6) had a mean age of 62.9 ± 13.1 years and a mean body-mass-index (BMI) of 27.1 ± 4.8 kg/m2 (see 2) and 12 samples from patients with a diagnosed GHD . The patients were separated by their type of GH-related disorder  and further divided into preoperative and postoperative, respectively. Nine newly diagnosed patients with acromegaly and five with a newly diagnosed GHD, who had not received any treatment yet, were assigned to the preoperative subgroups. The patients that donated blood to the study after their tumor surgery were assigned to the postoperative subgroups. The postoperative acromegaly group (n = 23) contained 15 samples with GH suppressive medication and eight samples that did not receive any medication. For postoperative GHD (n = 7), four samples were from patients who received daily GH replacement therapy while three did not receive treatment. All postoperative patients had a normal IGF-1 and therefore a controlled disease. A significant difference in mean age between all patients and controls existed (p = 0.002), but mean BMI values did not differ significantly (p = 0.08).The control group cohort (g/m2 see . The grop < 0.0001). Although their samples were within the clinical RR, the postoperative acromegaly group showed a significant increase in IGF-1 values vs. controls  but only the five preoperative GHD patients fell below the clinical RR. Postoperative GHD patients showed normal values, which were close to the lower reference range boundary but still lower than in controls . All preoperative acromegaly patients had IGF-1 levels above the clinical RR. The values were highly elevated in comparison to the control group , cullin-5  and Rbx-2  compared to controls. Only EloB showed no significant difference to controls . In contrast, most of these differences were not observed comparing the postoperative acromegaly group and controls (p = 0.001). Interestingly, the highest measured values of the four proteins derive from a patient, who was with 260 ng/mL IGF-1 barely above the clinical RR. In preoperative GHD patients, only EloB was lower than in controls , cullin-5 (p = 0.01) and Rbx-2 (p = 0.001) could be found in postoperative GHD samples compared to control samples. Although there was a significant difference in mean age between patients and controls, there was no linear dependency on age in SOCS2 , EloB , cullin-5  or Rbx-2 . Moreover, BMI values, which ranged from 18 to 35 kg/m2 in controls, also did not correlate with measured SOCS2 , EloB , cullin-5  or Rbx-2  levels. Furthermore, no significant gender differences in mean SOCS2 (p = 0.54), EloB (p = 0.13), cullin-5 (p = 0.43) or Rbx-2 (p = 0.35) protein concentrations were observed in controls. By clustering the results of all samples, the scaled combination of SOCS2, cullin-5 and Rbx-2 formed three clusters with all control samples falling into one cluster . Receiver operating characteristic (ROC) curves have been compiled to calculate area under the curve (AUC) values and to investigate the trade-off between sensitivity (SEN) and specificity (SPEC).For preoperative acromegaly A, SOCS2,In all four groups, the combined SCR index correctly classified all patients and controls in all tests. Furthermore, its SEN and SPEC was always 1.00 and the AUCs of the SCR index further substantiate its excellent diagnostic accuracy, as they range from 0.97 to 1.00 see A–D.This study is the first to investigate changes of all ECS-complex proteins in blood samples of patients with treatment-naïve or controlled GH disorders. Our data provide evidence for a correlation of these disorders with abnormal concentrations of ECS-complex proteins. While protein concentrations were significantly elevated in preoperative acromegaly, they were found to occur both elevated or reduced in GHD. The reasons for these discrepancies are not understood yet, as they could not be attributed to certain characteristics or therapy states of individual patients.In order to conform to the typical age distribution of patients with GH-related disorders, only controls above 40 years of age were recruited for the study. This resulted in a significantly higher mean age in controls vs. patients but did not seem problematic as none of the proteins indicated an age-related correlation. Problematic, however, is the broad reference range of the standard biomarker IGF-1, because it may complicate reliable diagnostic assessments as well as treatment control. In contrast, the ECS-complex proteins show a stable physiological expression due to small coefficients of variation which suggests narrow reference ranges. This would allow more precise diagnostics and furthermore may explain the measured abnormal ECS-complex concentrations in postoperative patients with normalized IGF-1 levels. Accordingly, these values could indicate that many patients may still harbor residual disease activity. This assumption is supported by research showing that the IGF-1 of patients with acromegaly can normalize with symptoms still in place . Similarp = 0.008). As this age-dependent effect could not be observed in controls, it may only apply to the stimulation of SOCS2 without affecting its basal levels. It further indicates that the SOCS2-dependent feedback loop is more active in younger patients, while it weakens in the elderly. Ultimately, this could impede the diagnostic capability of SOCS2 in older patients. In mice, SOCS2 was found to only carry out its inhibitory effect on GH signalling in lower concentrations, while it potentiated signalling at higher concentrations [Furthermore, no correlations between age, gender or BMI and the ECS-complex proteins in the control group samples have been found. Therefore, these biometric data may not have to be considered when using these proteins as biomarkers. It may also mean that the known gender-related effects of estrogen on the GH/IGF-1 axis may not affect the ECS-complex . This altrations ,54. AccoThe statistical capacity of the ECS-complex proteins as diagnostic biomarkers was estimated by comparing their performance parameters. Generally, the SPEC of all proteins was 100% in nearly all tests, which is probably due to the low variation of measured ECS-complex protein concentrations in controls. Only for cullin-5 in the preoperative groups, the calculated SPEC was lower (93%).Of the individual proteins, SOCS2 showed the best performance on average, as its diagnostic accuracy was over 0.9 in all four groups. Its advantage over the other ECS-complex proteins might be its direct stimulation by GH. Therefore, it may be of good use as a single biomarker for both acromegaly and GHD. However, clustering of the results revealed that a combination of SOCS2, cullin-5 and Rbx-2 yielded a better separation of patients and controls than any of the individual proteins. This accounted for all four subgroups including preoperative GHD, where EloB showed the highest significance but worsened the group separation. In fact, by using the SCR index, the SEN and overall diagnostic performance was further improved. The SCR index was able to correctly identify every patient as positive and every control as negative, therefore presenting perfect SEN and SPEC in all four groups. It also identified patients with a normal IGF-1 as positive, while rejecting all controls. On the contrary, studies assessing the value of IGF-1’s diagnostic capabilities for these diseases are controversial. A meta-analysis determined IGF-1’s SEN (0.66), SPEC (0.69) and AUC (0.78) for the diagnosis of a GHD, which indicates only moderate performance . BecauseLastly, there are some limitations in our study that should be pointed out. The comparison of the here investigated biomarkers to IGF-1 is biased because it was part of the initial diagnostic assessment for the patients in this study. Nonetheless, it can be added that the absence of correlation between any of the ECS-complex proteins and IGF-1 was expected as SOCS2-deficient mice do not show elevated IGF-1 expression after GH treatment . For a bThis is the first study to evaluate the potential of the ECS-complex for the diagnostics of GH-related diseases. We have demonstrated significant differences in all four investigated ECS-complex proteins between patients with acromegaly or GHD and healthy controls. The stable expression of the proteins in controls together with the significant differences in patients make up good characteristics for them to be used as alternative biomarkers. Measured protein concentrations also did not change in relation to age, gender or BMI, indicating independence from these factors.Overall, SOCS2 showed the best clinical performance in terms of SEN, SPEC and accuracy as an individual marker. However, superior results were obtained with a combination of SOCS2, cullin-5 and Rbx-2, which showed excellent diagnostic performance by reliably separating all patients from controls.Therefore, this combination of proteins is proposed as a novel biomarker candidate for acromegaly as well as GHD and because of its high sensitivity can be considered for the screening of such rare diseases as well. In the future, it may be an alternative for contradictory IGF-1 results, postoperative treatment control or hazardous provocative tests.


In addition, the proposed GO-based electrochemical sensor was tested using actual pharmaceutical naproxen tablets without pretreatments, further demonstrating excellent sensitivity and selectivity. Moreover, this study provided insights into the participatory catalytic roles of the oxygen functional groups of the GO-based nanomaterials toward the electrochemical oxidation and sensing of naproxen.Here we report on a selective and sensitive graphene-oxide-based electrochemical sensor for the detection of naproxen. The effects of doping and oxygen content of various graphene oxide (GO)-based nanomaterials on their respective electrochemical behaviors were investigated and rationalized. The synthesized GO and GO-based nanomaterials were characterized using a field-emission scanning electron microscope, while the associated amounts of the dopant heteroatoms and oxygen were quantified using x-ray photoelectron spectroscopy. The electrochemical behaviors of the GO, fluorine-doped graphene oxide (F-GO), boron-doped partially reduced graphene oxide (B-rGO), nitrogen-doped partially reduced graphene oxide (N-rGO), and thermally reduced graphene oxide (TrGO) were studied and compared via cyclic voltammetry (CV) and differential pulse voltammetry (DPV). It was found that GO exhibited the highest signal for the electrochemical detection of naproxen when compared with the other GO-based nanomaterials explored in the present study. This was primarily due to the presence of the additional oxygen content in the GO, which facilitated the catalytic oxidation of naproxen. The GO-based electrochemical sensor exhibited a wide linear range (10 µM–1 mM), a high sensitivity (0.60 µAµM Naproxen  propanoic acid (S/R)) is a nonsteroidal anti-inflammatory drug (NSAID) that is used to treat inflammation, fever, rheumatoid arthritis, and stiffness. Naproxen inhibits COX-1 and COX-2 enzymes, which results in the inhibition of the synthesis of certain prostaglandins ,4,5. CasMethods such as high-performance liquid chromatography (HPLC), UV-spectrophotometry, spectrofluorimetry, and mass spectrometry have been commonly employed to analyze pharmaceutical drugs; however, these techniques are generally time-consuming and costly. The electrochemical approach provides an accurate, fast, and cost-effective alternative to those detection methods . For insThe reduction of GO can cause significant structural defects, which may play important roles in catalysis ,20. HoweThe purpose of this study was three-fold: (i) to develop an advanced electrochemical sensor for the detection of naproxen; (ii) to compare the performance of various GO-based sensors and rationalize the effects of oxygenated functional groups; and (iii) to investigate the effects of dopants on the performance of the GO-based electrochemical sensors.2HPO4 and NaH2PO4, and NaOH was employed to adjust its pH to 7.2. GO was synthesized from high-purity graphite provided by ZEN Graphene Solutions Ltd., Thunder Bay, Canada, using a modified Hummers method  in 0.2 M KNO3) to compare their electrochemical performance. As seen in The modified electrodes  and pristine GCE were examined in a KNOThe CV curves recorded in a 0.1M PBS buffer (pH 7.2) in the absence (dashed line) and in the presence of 300 µM naproxen are presented in the 2 at 1.13 V. In contrast, the GO/GCE generated the second lowest current density (188.00 µA/cm2) and the second largest peak separation (121.0 mV) during the ferricyanide test  was the main contributor to the significant differences in the catalytic oxidation performance of the modified GO electrodes. For instance, the GO/GCE possessed 30.89% of the oxygen content compared to 15.39% for the TrGO. The rationale behind the performance of these electrodes may be that as the oxygen content of graphene becomes higher, the catalytic oxidation of naproxen becomes stronger. The only outlier was the F-GO/GCE, as it accounted for only 28.33% of the GO detection signal. However, it is known that fluorine doping completely disrupts the sp2 carbon ring structure, thus further decreasing the conductivity of GO [ide test a. Similaty of GO . This chty of GO .The robust adsorbing behavior of naproxen is primarily facilitated by the binding interactions between the graphene sp2 carbon rings and carboxylic acid groups of naproxen . Previou2 value of 0.9963, which signified a very strong linear relationship. The sensitivity of the sensor was obtained from the slope of the regression line to be 0.60 µAµM−1cm−2. The limit of detection (LOD) was calculated to be 1.94 µM, which was acquired using the formula LOD = 3 σ/s. The limit of quantification (LOQ) was calculated to be 6.47 µM, which was obtained using the formula LOQ = 10 σ/s, where σ represents the standard deviation of the five blank measurements, and s denotes the slope from the calibration curve.The selectivity of the developed GO/GCE sensor was also investigated; The performance of the GO/GCE sensor was further tested using a Life Brand Naproxen tablet (220 mg) as shown in In summary, five different GO-based nanomaterials including GO, TrGO, B-rGO, N-rGO and F-GO were synthesized and systemically studied. GO/GCE exhibited the strongest activity toward the electrochemical oxidation of naproxen compared to GCE and other modified GO electrodes. This was primarily due to an enhanced catalytic activity facilitated by the oxygen functional groups of GO, particularly the epoxy and hydroxyl groups, which was confirmed by the XPS analysis. The sensitive quantification of naproxen was successfully achieved over a wide linear concentration range from 10 µM to 1 mM by DPV. Naproxen had very strong propensity for fouling electrode surfaces, resulting in a substantial decrease of the current. A facile electrochemical activation strategy based on potential pulses was developed, which successfully overcame the critical fouling problem. The GO/GCE sensor developed in this study exhibited high sensitivity and selectivity over a wide range of concentrations of naproxen. Off-the-shelf naproxen tablets were successfully detected using the developed sensor, which demonstrated strong anti-interference capabilities. Future studies should be conducted to elucidate the specific naproxen catalytic kinetics with oxygenated functional groups, as well as heteroatom dopant levels, catalytic oxidation and antifouling properties toward naproxen.